DEPARTMENT OF LIFE SCIENCES

FACULTY OF SCIENCE AND TECHNOLOGY

BIOL2764
PHYSIOLOGY OF PLANTS

LABORATORY MANUAL
2017 – 2018
TABLE OF CONTENTS

Introduction

Practical 1 - Water Relations

1.1 Estimation of the water potential of potato tubers by length changes

1.2 Determination of the solute potential of cell sap
1.3 Plant water potential measured with the Pressure Chamber

Practical 2 – Photosynthesis and gas exchange

2.1 Measurement of leaf diffusive resistance with the Porometer

2.2 Measuring photosynthesis using an infra-red gas analyzer (IRGA)
2.3 Effect of light level and herbicides on photosynthesis of leaf discs

Practical 3 – Germination, Mineral Nutrition and Hormone Response

(Week A: preparation)

3.1 The role of light and hormones in germination of light sensitive seeds
3.2 Hormones and growth analysis
3.3 Hormones and apical dominance
3.4 Commercial use of hormones
3.5 Description of deficiency symptoms for selected mineral nutrients

Practical 3 – (Week B: Interim measurements)

Practical 4 – (Week C: results)

Introduction

Requirements for write-up and submission of Labs

You are required to print the relevant section of the Lab manual before the practical. You should
read over the manual and the associated class material so you are ready to submit the
completed report at the end of the practical. All practical reports are to be submitted at the
end of the practical, except practical 3 which is submitted at the end of practical 4.

Marked reports will be distributed in the subsequent practical. The penalty for late submission of
books is as follows: 10% deducted per day late; no acceptance of books after 7 days.

Coursework components
The coursework component of BIOL2764 carries 50% of the total course mark. The coursework
comprises of four Laboratory Practical reports and two In-course Examinations. The allocation
of marks for these components is as follows:

 Laboratory Practicals 20 %

 In-course Examination I 15 %

 In-course Examination II 15 %

Laboratory Safety and Etiquette

 Lab coats, closed shoes and long trousers/skirts must be worn at all times in the lab.

 No eating, drinking, smoking, chewing gum, or applying cosmetics in the laboratory.

 Always wash your hands before leaving lab. It is a good idea to wash your hands at
intervals during your experiments.

 Clean up your work space and materials at the end of your experiment.

 Report all accidents and breakages, no matter how small, to your demonstrator. Broken
glass and sharp objects must be disposed of in the ‘sharps’ bin provided.

 Learn the location of all safety equipment including first-aid kit, fire extinguisher, safety
shower, eyewash station, and acid/alkali spill cleanup materials.

 Be aware of the chemicals you will be using. Using an online Google search
(www.google.com) copies of MSDS for any chemical can be obtained.
Practical 1 - Water Relations

1.1 Estimation of the water potential of potato tubers by length changes

1.2 Determination of the solute potential of cell sap
1.3 Plant water potential measured with the Pressure Chamber

Introduction
The main factors influencing water potential in plant cells can be summarized using the water potential
equation:

Ψw = Ψs + Ψp

Where, Ψw = water potential, Ψs = solute potential, Ψp = hydrostatic pressure

The solute potential of most solutions can be estimated using van’t Hoff’s equation:
Ψs = -MiRT (MPa)

M = molal concentration of solution (mol kg-1)

i = ionization constant (i = 1 for sucrose)
R = gas constant (0.00831 kg MPa mol-1 K-1)
T = Absolute temperature (K) (to convert from C to K, add 273)
For our Practical Sessions, assume that Molarity  Molality (this is so for dilute solutions).

In these experiments we will be using sucrose solutions. Sucrose is small enough to pass through the cell
wall but too large to pass through the plasma membrane by simple diffusion (sucrose can be
transported into a cell by transport proteins through facilitated diffusion).

1.1 Estimation of the water potential of potato tubers by length changes

Using the 1 M sucrose stock solution provided and a 250 beaker, make up 100 ml of each of the
following concentrations: 0.05, 0.10, 0.15, 0.20, 0.25 and 0.30 M (these will be used for experiment 1.1
and experiment 1.2). Pour each solution into labelled petri-dishes with a lid, use enough solution to fill
the dish.

Select a large, firm potato (Solanum tuberosum) tuber and cut out 12 cylinders using a cork borer (5-6
mm diameter). Do not push the cork borer directly onto the bench surface, use the cardboard provided to
protect the bench. Trim each cylinder to 40 mm length with a razorblade, making sure that all suberized
layers are removed. Place each cylinder immediately into a small beaker that is kept covered to
minimize evaporation from the cut surfaces until all cylinders are cut. After recording the exact lengths
to the nearest 0.5 mm, place 2 cylinders (randomly selected) into each of the sucrose solutions. Note
the time and cover each petri-dish to prevent evaporation.

After an equilibration time of 60 min, remove the potato cylinders from each of the solutions and blot
lightly on a paper towel. Quickly determine the new length of each cylinder to the nearest 0.5 mm.
Table 1.1. Initial length of potato cylinders

Length (mm) Sucrose concentration

0.05 M 0.10 M 0.15 M 0.20 M 0.25 M 0.30 M

Cylinder #1

Cylinder #2

Average

Table 1.2. Final length of potato cylinders (after 60 minutes) and the percentage change

Length (mm) Sucrose concentration

0.05 M 0.10 M 0.15 M 0.20 M 0.25 M 0.30 M

Cylinder #1

Cylinder #2

Average

% Change

(Use +/- to indicate increase/decrease in average length of cylinders)

Using the grid on the next page, plot a graph of the percent change in length against the concentration
of the sucrose solution, and draw a straight line that best fits the data points.

From your graph, what sucrose concentration results in zero % change in length of tissue?

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100
90
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70
60
50
40
30
20
10

-10
-20
-30
-40
-50
-60
-70
-80
-90
-100
Calculate the solute potential of the sucrose solution that resulted in zero change in length (using van’t
Hoff’s equation). For sucrose solution the solute potential is equal to the water potential. Explain why?

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The water potential of the potato tissue is equal to that of the sucrose solution at which there was zero
change in length. Explain why.

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The plant cell is enclosed by a selectively permeable membrane. How would the results have changed if
the membrane was fully permeable to sucrose?

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1.2 Determination of the solute potential of cell sap
From the 1.0M sucrose stock solution provided, prepare 10 ml of each of the following solutions:
0.05M, 0.1M, 0.15M, 0.2M, 0.25M, 0.3M and 0.35M sucrose.

Remove 7 epidermal peels from the lower epidermis of Rhoeo (Tradescantia spathacea) and place them
in distilled water for about 5 min. Transfer 1 strip to each of the sucrose solutions and ensure that strips
are completely immersed in the solutions. After 15 mins, examine the strips under a microscope by
mounting each strip in its bathing medium on the slide. Count the number of plasmolysed and
unplasmolysed cells within three randomly selected fields-of-view (FOV). Restrict your observations to
anthocyanin-pigmented cells (record your results in Table 1.3).

Plot % plasmolysed cells vs. sucrose concentration, and draw a straight line that best fits the data points.

100
90
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30
20
10

From your graph, what sucrose concentration results in 50% plasmolysis?

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Table 1.3 Number of plasmolysed and unplasmolysed cells, and % Plasmolysis

Sucrose Number of cells Number of cells % Plasmolysis Average %

conc. Plasmolysed Unplasmolysed Plasmolysis
FOV

0.05 M #1

#2

#3

0.10 M #1

#2

#3

0.15 M #1

#2

#3

0.20 M #1

#2

#3

0.25 M #1

#2

#3

0.30 M #1

#2

#3

0.35 M #1

#2

#3
Calculate the water potential of this sucrose solution from van’t Hoff’s equation. When 50% of the cells
reach plasmolysis the tissue has reached ‘incipient plasmolyses’. The water potential that results in
incipient plasmolysis is equal to the solute potential of the tissue at incipient plasmolysis. Explain how
this experiment determines the tissue solute potential rather than tissue water potential.

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1.3 Plant water potential measured with the Pressure Chamber
In order to transport water up to the top of the canopy, plants must maintain a gradient of water
potential from the roots to the upper leaves. This is sometimes referred to as the soil plant atmosphere
continuum (SPAC). Here, we will measure the water potential of leaves from the lower and upper
branches of a tree from the UWI campus. Taking all precautions necessary, use the Pressure Chamber
method to measure leaf water potential.

Method
A leaf is excised with a single clean cut, and then sealed into a chamber with the cut end of the petiole
protruding through a rubber seal. The excision releases the tension (negative hydrostatic pressure) in
the xylem causing sap to move from the xylem into the surrounding tissue. Inside the chamber the
pressure is increased slowly until the xylem sap just begins to exude from the cut end of the plant tissue.
Pressurization is stopped at this point and the air pressure in the chamber recorded. The pressure that is
required to force the xylem sap back to the cut surface is approximately equal to the original hydrostatic
pressure of the xylem. As solute potential of the xylem is usually negligible (i.e. the xylem sap is very
dilute), this is also an estimate of xylem water potential.

In a non-transpiring leaf the water potential of the xylem and the water potential of the surrounding
tissue are at equilibrium, so this also serves as an estimate of leaf water potential.

Precautions:

 Evaporative loss from the leaf should be prevented. Enclosing and sealing the sample in a
humidified plastic bag is recommended. When there is no transpiration, the water potential of
the leaf cells and the water potential in the xylem will come into equilibrium.

 Tension on the seal should be just sufficient to prevent leakage. However, the walls of xylem
vessels are usually very resistant to constriction.

 Increase pressure slowly (0.05 - 0.2 bar s-1), and do not over-pressurize. Sap is lost if tissues are
over-pressurized and balance pressures will be too high.

Table 1.4. Leaf water potential values (MPa).

1 2 3 4 Mean

Upper

Lower

Difference

*note: 1.0 Bar = 0.1 MPa

*note: water potential will have the opposite sign (+/-) to balancing pressure).
What difference do we expect in the water potential of upper and lower leaves? Did you detect this
difference, if you didn’t suggest reasons for this?

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How would you expect the results to change if the plants were under water stress?

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Practical 2 – Photosynthesis and gas exchange

2.1 Measurement of leaf diffusive resistance with the Porometer

2.2 Measuring photosynthesis using an infra-red gas analyzer (IRGA)
2.3 Effect of light level and herbicides on photosynthesis of leaf discs

Introduction
Last week we looked at the processes that determine the uptake of water from the soil. Here we look at
the processes that control the release of water from the leaves to the atmosphere (transpiration) and
the exchange of gases during photosynthesis and respiration (gas exchange).

2.1 Measurement of leaf diffusive resistance with the Porometer

For a well watered plant the transpiration rate is determined by the atmospheric conditions around the
leaf and by leaf conductance. Leaf conductance determines the rate of water loss from the leaf surface
under different atmospheric conditions. It is measured in mmol m2 s-1, i.e. how many mmol of water
cross a m2 of leaf surface in one second. The higher the conductance the more transpiration will occur.
The inverse of leaf conductance is leaf diffusive resistance (m2 s mmol -1).

Calibrate the automatic diffusion porometer, and measure the leaf conductance on the lower (abaxial)
and upper (adaxial) leaf surfaces for 2 plant species provided (Sugarcane and Cocoa).

Procedure for Measuring Leaf Stomatal Conductance:

Leaf conductance can be measured using a diffusion porometer. You will use the Decagon Porometer
(SC-1). The instrument consists of a small chamber that is clamped to the leaf surface, inside of the
chamber are two humidity sensors at different distances from the leaf. Water evaporating from the leaf
through the stomata causes the humidity measured by the two sensors to increase, the rate at which
this happens is used to calculate the rate of transpiration from the leaf surface.

Calibration:
Each time the porometer is used it must be calibrated (this will be done before the Practical by the
technicians). The porometer is calibrated by measuring the rate of evaporation from filter paper wetted
with deionised water. The filter paper is covered by perforated polypropylene plates with known pore
sizes. Plates with large pore sizes results in a higher rate of evaporation. This is repeated with several
plates each with a different pore size and the results are used to create a calibration equation which is
stored by the machine.

Precautions:
Measurements are affected by the temperature of the chamber. Try not to expose the chamber head to
any changes in temperature (e.g. direct sunlight, or touching the metal part of the chamber). Ensure a
proper seal between the leaf surface and the porometer chamber. Leakage of air around the edge of
the chamber can lead to variable readings and errors.

Make sure you know which side of the leaf is begin measure for every measurement (Abaxial-lower
surface or Adaxial-upper surface).
Table 2.1. Porometer measurements for each plant species

Species Leaf Leaf conductance (mmol m2 s-1)

surface
Rep #1 Rep #2 Rep #3 Rep #4 Mean

Species #1 Abaxial

Adaxial

Ratio

Combined

Species #2 Abaxial

Adaxial

Ratio

Combined

What aspect of the leaf anatomy will influence leaf conductance?

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Is there difference in the conductance of upper and lower leaf surfaces. What is the physiological
purpose of the difference?

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Describe any differences in leaf conductance between the species

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2.2 Measuring photosynthesis using an infra-red gas analyzer (IRGA)

With the assistance of the laboratory technician obtain net CO2 exchange rates for leaves of one of the
three plant species provided using the open gas exchange system (Successive groups will measure a
different species and the results will be shared).

A portable gas exchange system will be used to measure net CO2 exchange rates. The system consists of
an air supply unit, an infrared gas analyser, a leaf chamber and a data logger. This gas exchange system
is described as being an open-system in that air is drawn from the atmosphere (4m above), flows
through the instrument, and returns to the atmosphere. Other systems are also available where the
same air is re-circulated to the chamber throughout the measurement (closed systems).

The operation of the infrared gas analyser (IRGA) is based on the principle that hetero-atomic gas
molecules (such as H2O & CO2) absorb irradiation at specific infrared wavelengths. Air from the air
supply unit (300 mL min-1) is split into two streams. The ‘reference’ stream flows directly to the IRGA
and indicates the CO2 concentration of the incoming air, while the ‘analysis’ stream flows first to the leaf
chamber and then to the IRGA. The photosynthetic rate is calculated from the difference in CO2
concentration between the ‘reference’ and ‘analysis’ streams. Sensors inside the leaf chamber measure
leaf and air temperature, light (photosynthetic photon flux density (PPFD)) and relative humidity. After
enclosing the leaf in the leaf chamber steady-state readings can be reached within 60 s, and net
photosynthesis (mol CO2 m2 s-1) can be measured.

Table 2.2 Obtain information on the gas exchange system and measurement conditions.

Air flow rate into the chamber

Projected leaf area exposed in the chamber

Atmospheric temperature

Irradiance reaching the leaf chamber

Table 2.3 For one of the plant species provided, record the CO2 concentration entering (CO2 –in) and
leaving (CO2 –out) the leaf chamber (as parts per million) and the net CO2 exchange rate (in μmol m2 s-1).

Plant species CO2 – in CO2 – out CO2 exchange

rate

Explain why the parameter measured is net CO2 exchange rate rather than gross CO2 exchange rate?

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Light response curve:
The aim of this experiment is to produce a light response curve. This curve shows how the
photosynthetic rate increases with light intensity. This curve gives a lot of information about the
photosynthetic capacity of the plant. For each of the plant species provided, use the graph produced by
the IRGA to estimate the following parameters: light compensation point, dark respiration rate and light
saturation point.

Species #1 Light compensation point:

Dark respiration rate:

Light saturation point:

Species #2 Light compensation point:

Dark respiration rate:

Light saturation point:

Describe the components of the curve- light compensation point?

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Describe the components of the curve- light saturation point?

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Describe the differences in gas exchange for the two species used:

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2.3 Effect of light level and herbicides on photosynthesis of leaf discs

Here we measure photosynthesis in leaf discs by looking at gas exchange in a solution. We will produce
a light response curve and investigate the impact of herbicides on the response.
Herbicides work by disrupting essential metabolic processes within plants. In this experiment we will
investigate the effect of two herbicides on the light response:
Diuron (DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea)) works by blocking the plastoquinone binding site of
photosystem II.
Gramoxone (Paraquat) works by reacting with oxygen inside the chloroplasts to form damaging free radicals,
which result in lipid peroxidation.
Your results should help you understand the impact these herbicides have on the plant.

Avoiding the major veins, punch out about 100 leaf discs from the Amaranthus plants provided. Float
the leaf discs on Sodium Bicarbonate solution (NaHCO3, 1%) under low light conditions for about half-
hour, then infiltrate the discs with the same Sodium Bicarbonate solution by applying suction with a
syringe (actual procedure will be demonstrated during the lab session). Repeat the indicated procedure
until most or all discs sink to the bottom of the solution. Discard any leaf discs that do not sink, and
keep the Sodium Bicarbonate-infiltrated discs away from light. These Sodium Bicarbonate-infiltrated
discs, sink as the air in their intercellular spaces has now been replace by the denser Sodium Bicarbonate
solution.
For each treatment, remove 5 leaf discs and place them in a small petri-dish containing 25mL of Sodium
Bicarbonate solution. Place the petri-dish so that the light reaching the leaf discs is mainly from a light
source of known intensity. Set the distance of the light source from the petri-dish, turn on the light
source and note the time taken for each leaf disc to rise to the surface. Vary the distance (10, 20, 30, 40
cm) of the light source from the petri-dish using fresh samples of leaf discs for each distance. Measure
the light intensity at each distance using a light meter.

Use this experimental procedure to study the effects of the two herbicides provided on rates of
photosynthesis. To apply the herbicide treatment use 2 drops of herbicide per petri-dish and leave for
15 min prior to light exposure (Ask your demonstrator before handling these compounds as they are
harmful if inhaled or through skin contact).
Control Leaf disc # 10cm 20cm 30cm 40cm

Mean

Diuron Leaf disc # 10cm 20cm 30cm 40cm

Mean

Gramoxone Leaf disc # 10cm 20cm 30cm 40cm

Mean
For each treatment, plot the reciprocal of the mean time for leaf discs to float vs. the measured light
level and fit a straight line by eye.
 Explain the response to light of the control discs (why do the discs float; what happens when the light is
increased?).

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Explain the response to light of the herbicide treated discs and suggest reasons for differences between
the treated and the control discs.

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Practical 3 & 4 – Germination, Mineral Nutrition and Hormone Response

Introduction
The following experiments will run over three weeks as outlined in the Table below. This includes an
interim week between the practicals. The results from all the experiments will be submitted at the end
of practical 4.

WEEK A WEEK B WEEK C

3.1 The role of light and hormones in prepare measure submit

germination of light sensitive seeds

3.2 Hormones and growth analysis measure measure complete

/submit

3.3 Hormones and apical dominance complete submit

3.4 Commercial use of hormones complete submit

3.5 Description of deficiency symptoms for measure measure complete

selected mineral nutrients /submit

Week B Interim measurements

After Practical 3 each group must arrange with the Teaching Assistant to make interim measurements of
the following:

After two days: Germination of light sensitive seeds (this experiment will be discontinued after two days).
You must submit your results to the Teaching Assistant. Data from all of the groups will be available from
Myelearning. You must download the data, carry out a statistical analysis and write a scientific abstract.

After one week: Hormones and growth analysis. Harvest the plants from treatments 2&3 and place in
the oven. Use the same procedure as for treatment 1 (including the measurements of leaf area) and
clearly label all material with your group number.

After one week: Mineral nutrition. Interim observations of nutrient deficiency are necessary to
distinguish between the different treatments.

In your own time: Commercial use of hormones. You will need to compile this information before the next
practical.
3.1 The role of light and hormones in germination of light sensitive seeds
Seeds of lettuce (Lactuca sativa; variety Grand Rapids) are dormant when freshly shed. This dormancy
can be broken when seeds are hydrated (imbibition) and exposed to light. This light affect is mediated
by the pigment-protein phytochrome, which is synthesized in the mature dormant seed as the red
absorbing form (Pr). The Pr form undergoes a conversion to the far-red absorbing form (Pfr) upon
exposure to red light. Conversion of Pr to Pfr by exposure to red light promotes germination. Certain
hormones also affect seed germination by reducing or eliminating the light requirement.

Week A:
Place a piece of filter paper in each of five small petri dishes. Identify treatments by labelling the top of
the petri dishes in a way which will not block light from the seeds. Spread 20 seeds over the filter paper
in each petri dish. Add sufficient distilled H2O to moisten the filter paper in each petri dish, except in
treatment 3 where GA3 solution is added instead. Seal the edges of all the petri dishes with plastic wrap
to prevent desiccation of developing seedlings. For treatment 2 & 3, wrap dishes with 2 layers of
aluminum foil. Treatments 4 & 5 will be placed in boxes with coloured cellophane filters that block the
transmittance of light at specific wavelengths. All work must be done in the dark room using a green
save light.

Approx. 20 seeds will be added to each.

Treatment Description
1 White light
2 Dark (control)
3 Dark + 50 ppm GA3
4 Far-red light
5 Red light

Week B & C:
After two days remove the lids and count the number of germinated seeds in each dish. It is important
that you do not disturb other student’s seeds or expose them to light at this time. Record the
percentage germination and submit the data to the Teaching Assistant. The data from all the groups will
be distributed on Myelearning. Each person must carry out a statistical analysis of the data (using
ANOVA), and write an abstract on the effects of gibberellic acid and light on germination of the lettuce
seeds.
 Germination %

Treatment Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Mean

Results from ANOVA

Your abstract should follow the format used in scientific journals. The general outline is given in the
template below (only 1 or 2 short statements are needed for each heading).

Introduction (or rationale for the study):

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Materials and Methods:

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Results:
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Discussion:
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Conclusion:
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3.2 Hormones and growth analysis
Here we use classical growth analysis to investigate the impact of hormones on plant development. We
will use many of the formulas discussed in the Lectures:

Absolute growth rate = (dW/dT) = (W2-W1) / (T2- T1) [g day-1]

Relative growth rate = (dW/dT) x (1/W) = (W2-W1) / (T2- T1) x (1/W1) [g g-1 day-1] %

Net Assimilation Rate

NAR = (dW/dT) x (1/leaf area) = (W2-W1) / (T2- T1) x (1/ leaf area) [g day-1 m-2 ]

Leaf Area Ratio

LAR = (leaf area / plant weight (W)) [m-2 g-1 ]

Leaf Area Index = leaf area / ground area [m-2 m-2 ratio units]

Specific Leaf Area

SLA = (leaf area / leaf weight) [m-2 g-1 ]

Root: Shoot ratio

(root weight / shoot weight) [g : g ]
Week A:
Investigate the effects of gibberellic acid (GA) spray on the growth of bodi (Vigna unguiculata) seedlings.
You are provided with one-week-old bodi seedlings growing in seedling trays. Each group should select
three small trays containing seedlings. Randomly assign one of the following treatments to each tray:

1. Initial 2. Control 3. GA treated

For ‘3 GA treated’, plants are to be sprayed with 100ppm gibberellic acid (GA3) to wet all leaves (ask
your demonstrator for help with this). Plants are to be left unsprayed in treatments 1 and 2.

Treatment 1, this will be used to estimate the biomass of the plants at the start of the experiment and
provide a reference for the growth that will occur in the two other treatments. Carefully cut the stem of
the plant at the soil level:

 Divide each plant into (a) stem plus petioles (b) leaf.

 For leaf area index, measure the total leaf area per plant. To estimate leaf area, draw outlines of
the leaves on graph paper then cut out the shapes and weigh them. A separate section of graph
paper of known area is also weighed to give a conversion factor for calculating leaf area.
You will also need an estimate of the ground area occupied by each plant- this is approx. 10 cm2.

 For biomass (dry weight), combine the 5 plant samples and place stems and leaves separately
into labelled paper bags and dry at 80°C to constant mass (you need only 2 paper bags). Dry
mass will be taken after two weeks (note: constant mass will be achieved after 2-3 days, but the
material can stay in the oven until the next Lab session).

Week B:
Repeat all the above measurements for treatments 2 and 3 after one week of growth. Theses will be
kept in the oven until Week C.

Week C:
Weigh all samples from week A and Week B.
Leaf area conversion factor

Week A Week B

Mass (g) of 100 cm2 graph paper

Area conversion factor (cm2 per g paper)

Ground area (cm2) occupied per plant (avg. row × column spacing between plants)

Table 3.1 Results for treatment 1. Initial

Variable Bodi (initial)

Leaf area (cm2 per plant)

Leaf mass (g per plant)

Stem mass (g per plant)

Total above ground mass

(g per plant)

Table 3.2 Results for Control and GA treated plants

Control Bodi GA treated Bodi

Leaf area (cm2 per plant)

Leaf mass (g per plant)

Stem mass (g per plant)

Total above ground mass

(g per plant)
Table 3.3 Using the mean values above, calculate growth indices for your data (give units).
Measurements made at both times (weeks A & B) are used for calculation of AGR, RGR and NAR. Use
data only from week B to calculate the other indices.

Growth index Bodi

Control GA treated

Total mass

Leaf area

AGR

RGR

NAR

LAR

LAI

SLA
Give short statements to describe and/or explain the effects of the gibberellic acid treatment on each of
the following indices (from results given in Table 3.3)

Absolute growth rate

Relative growth rate

Net assimilation rate

Leaf area ratio

Leaf area index

Specific leaf area

Summarize the major effects of the GA treatment on growth of the plant species.

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3.3 Hormones and apical dominance
Tomato (Solanum lycopersicum) seedlings were decapitated (by cutting just below the apical bud) and
the hormone Auxin was applied to the cut stem as indole-3-acetic acid (IAA) in a paste. For control
plants an inert paste was applied to the cut surface without auxin. This experiment was set up on three
occasions: 4, 7 and 10 days ago. Make observations on the growth of axillary buds (if any) in these
plants.

Table 3.4 Length (mm) of the topmost axillary bud

Treatment applied 4 Treatment applied 7 Treatment applied 10

days ago days ago days ago
Replication
Paste IAA-Paste Paste IAA-Paste Paste IAA-Paste
alone alone alone

Mean

Give an physiological explanation for the difference you observed between the control and treated
plants:

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3.4. Commercial use of hormones
Week B&C:
Obtain details on the use of a plant growth regulator in any of the following:

 Control of fruit ripening in bananas or tomatoes

 Induction of flowering in pineapples

 Stimulation of rooting in cuttings

 Weed control

Select only one example and including product names, active ingredients, concentrations used and
application techniques (you can obtain information from: internet sources, visits to garden shops,
conversations with farmers, etc.).

Give details of the commercial use of a hormone/plant growth regulator.

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3.5 Description of deficiency symptoms for selected mineral nutrients

Week A, B & C:
A mineral is defined as essential if normal growth and reproduction cannot be carried out in its absence.
Plants usually respond to the deficiency of an essential mineral by characteristic visual symptoms and
stunted growth. Such symptoms are of interest because they shed light on the necessary function of the
element in the plant and because they are used by agriculturists to determine how and when to fertilize
their crops. A trained observer can predict the mineral deficiency of a given soil simply by examining the
conditions of the plants growing in it.

In this experiment you will record the development of symptoms in plants growing under several
mineral deficient conditions with coded labels. At the end of three weeks you will attempt to determine
which deficiency exists in each set of plants.

The roots of three healthy sand-grown seedlings were rinsed and placed into one of ten containers, each
of which was filled with one of ten aerated experimental solutions. Only the distilled water and the
complete medium treatments are labelled. The other five treatments (- Ca, - Mg, - N, - P, - Fe, and –
micronutrients) are coded.

Stock solution to add for each treatment (ml)

Stock solution Complete Ca Mg N P Fe Micro H2O

0.5M Ca(NO3)2 5 0 5 0 5 5 5 0

0.5M KNO3 5 5 5 0 5 5 5 0

0.5M MgSO4 2 2 0 2 2 2 2 0

0.5M KH2PO4 1 1 1 1 0 1 1 0

FeNaEDTA 1 1 1 1 1 0 1 0

Microelements 1 1 1 1 1 1 0 0

0.5M NaNO3 0 5 0 0 0 0 0 0

0.5M MgCl2 0 0 0 0 0 0 0 0

0.5M Na2SO4 0 0 2 0 0 0 0 0

0.5M NaH2PO4 0 0 0 0 0 0 0 0

0.5M CaCl4 0 0 0 5 0 0 0 0

0.5M KCl 0 0 0 5 1 0 0 0
For the next three weeks, make weekly observations of plant height, leaf number and visual appearance
(avoid touching the plants as this can cause damage). A list of typical deficiency symptoms is included
below for your guidance (Table 3.1). Symptoms to look for include:
Colour changes and their location; Dead spots and their location; Peculiarities of venation; other
pertinent information

In the tables below make your suggestion for which mineral is absent from each solution, indicate which
symptoms led you to make your choices. Be specific:

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Does the presence of a symptom on young and old leaves give you any information as to the mobility of
a each mineral? Cite specific examples from your data to support your answer.

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Person ……………………………………………………………………… Date………………………………………………………………….

Suggested
Observations mineral absent

Complete
1 minerals

Distilled water
8 (no minerals)
Person ……………………………………………………………………… Date………………………………………………………………….

Suggested
Observations mineral absent

Complete
1 minerals

Distilled water
8 (no minerals)
Person ……………………………………………………………………… Date………………………………………………………………….

Suggested
Observations mineral absent

Complete
1 minerals

Distilled water
8 (no minerals)
Table 3.1 Typical nutrient deficiency symptoms seen in plants (treatments included here are in bold)

Mineral Deficiency symptoms

Calcium Newer or upper leaves affected; young leaves of terminal bud typically hooked,
(Ca) later dying back at tips and margins. Growth characterized by a cut-out
appearance; stalk dies at the terminal bud; growth is restricted.

Magnesium Older or lower leaves affected; mottled or chlorotic leaves; leaves may redden;
(Mg) sometimes with necrotic spots; tips and margins turned upwards; stalk slender;
interveinal chlorosis with veins remaining green.

Nitrogen Older or lower leaves affected; plant light green; lower leaves yellow, drying to
(N) light brown colour; stalks short and slender if deficiency at later stage of growth;
a reddish or purple colour may occur along the veins.

Phosphorus Older or lower leaves affected; plant dark green, often developing red and
(P) purple colours; stalks short and slender if deficiency occurs in later stages of
growth; necrotic areas may develop on leaves, petioles and fruit.

Iron Newer or upper leaves affected; young leaves chlorotic, may become almost
(Fe) white; principal veins typically green; stalks short and slender.

Micronutrients Usually a delayed response with symptoms most evident in the youngest leaves.
Typical symptoms are: Manganese – spots of dead tissue on leaf, smallest veins
tend to remain green giving a reticulated pattern. Copper – young leaves
permanently wilted without spotting or marked chlorosis, twig or stalk below tip
unable to stand erect in later stages. Zinc – rapidly enlarging necrotic areas
between veins, leaves thick, shortened internodes. Boron – young leaves of
terminal bud light green at base, with final breakdown here, leaves become
twisted, stalk dies back at terminal bud. Molybdenum – required for nitrogen
metabolism, so symptoms resemble nitrogen deficiency.

Potassium Older or lower leaves affected; mottled or chlorotic leaves with spots of dead
tissue usually at tips and between veins; also seen on leaf margins; stalks
slender; plants stunted.

Sulphur Newer or upper leaves affected; young leaves with veins and tissue between
veins light green; growth continues normally; apical leaves appear quite yellow
to white.